Abstract: The study was aimed to evaluate the effect of Biofield Energy Treated vitamin D3 and DMEM medium on bone health parameters such as ALP, collagen, and bone mineralization in human bone osteosarcoma cells (MG-63). The test items (TI), were divided into two parts. One part of each sample received the Consciousness Energy Healing Treatment by Haddon Norman Salt and those samples were labeled as the Biofield Energy Treated (BT) samples, while the other parts of each sample were denoted as the untreated test items (UT). The cell viability by MTT assay data showed that the test samples were found as safe in the tested concentrations. The level of ALP was significantly increased by 73.24% and 85.41% in the UT-DMEM + BT-Test TI and BT-DMEM + BT-TI, respectively at 10 µg/mL compared to the UT-DMEM + UT-TI group. Further, ALP level was significantly elevated by 76.71% in the BT-DMEM + UT-TI group at 0.1 µg/mL compared to the untreated. Collagen was significantly increased by 77%, 113.53%, and 98.00% in the UT-DMEM + BT-TI, BT-DMEM + UT-TI, and BT-DMEM + BT-TI, respectively at 50 µg/mL compared to the untreated. Further, the collagen level was significantly increased by 128.05% and 132.10% in the UT-DMEM + BT-TI and BT-DMEM + BT-TI groups, respectively at 100 µg/mL; while increased by 76.31% in the BT-DMEM + UT-TI at 10 µg/mL compared to the untreated. Besides, the percent of bone mineralization was distinctly increased by 82.43%, 158.97%, and 52.95% at 0.1 µg/mL in the UT-DMEM + BT-TI, BT-DMEM + UT-TI, and BT-DMEM + BT-TI groups, respectively compared to the untreated. The percent of bone mineralization was significantly increased by 118.41%, 197.42%, and 116.94% in the UT-DMEM + BT-TI, BT-DMEM + UT-TI, and BT-DMEM + BT-TI, respectively at 1 µg/mL compared to the untreated. Further, bone mineralization was significantly increased by 202.34%, 200%, and 235.64% in the UT-DMEM + BT-TI, BT-DMEM + UT-TI, and BT-DMEM + BT-TI, respectively at 10 µg/mL than untreated. Altogether, the Biofield Energy Treated vitamin D3 was significantly improved the bone health parameters and it could be an excellent alternative nutraceutical supplement for vitamin D3 deficiency and fight against various bone-related disorders including osteoporosis, low bone density, osteogenesis imperfecta, Paget’s disease, rickets, osteomalacia, deformed bones, autoimmune and inflammatory diseases, stress management and prevention, and anti-aging by improving overall health.